Name: injection of an. stephensi embryos to generate malaria-resistant mosquitoes
Uploaded: 2007-07-04T00:00:00
Description: Watch this Scientific Journal Video about Injection of An. stephensi Embryos to Generate Malaria-resistant Mosquitoes at JoVE.com

<p class="jove_title"> Vorbereitung im Vorfeld der Mikroinjektion: </p><ol><li> Blut ernähren Mücken: für die Injektion von Montag bis Mittwoch Feed Weibchen auf der vorigen Freitag. Um Donnerstag und Freitag, Futtermittel Frauen am Montag injizieren der gleichen Woche. <br /><br /></li><li> Bereiten Sie den Quartz Nadeln mit dem Programm 2 und isotonischen Puffer (siehe Materialien). <br /><br /></li><li> &quot;Legen tube&quot; für die Embryonen ist die regelmäßige Drosophila Kultur Durchstechflasche. Wet Watte in den Boden, mit einem na...

<!DOCTYPE html PUBLIC "-//W3C//DTD XHTML 1.0 Transitional//EN" "http://www.w3.org/TR/xhtml1/DTD/xhtml1-transitional.dtd">
<html xmlns="http://www.w3.org/1999/xhtml" xml:lang="en" lang="en">	
		
		<div>
		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>Injection Quartz needles</td>
<td>&nbsp;</td>
<td>Sutter Instrument</td>
<td>&nbsp;</td>
<td>O.D.- 1.0 mm, I.D.-0.70, 10 cm length, are prepared by drawing capillary tubing into a fine 3-8 um with a shaft of approximately 100-300 mm micropipette puller. </td>
</tr>
<tr>
<td>Quartz puller</td>
<td>Tool</td>
<td>Sutter Instrument Co</td>
<td>P-2000</td>
<td>Set-up: Heat-275, Fil-3, Vel-38, Del-250, Pull-141</td>
</tr>
<tr>
<td>EndoFree plasmid kit</td>
<td>&nbsp;</td>
<td>Quiagen</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Isopropanol</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Millex-GV column</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>SLGV RO4 NL</td>
<td>for DNA cleaning</td>
</tr>
<tr>
<td>Construct DNA</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>0.5 mg/ml </td>
</tr>
<tr>
<td>Helper plasmid</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>0.3 mg/ml </td>
</tr>
<tr>
<td>Injection solution</td>
<td>Buffer</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>1x solution: 5mM KCl, 0.1mM sodium phosphate, ph 6. 8</td>
</tr>
<tr>
<td>Isotonic buffer</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>see recipe in the protocol section</td>
</tr>
<tr>
<td>Dessication oil</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Halocarbon oil 700:Halocarbon oil 27(1:1). Mix well. Prepare in advance of microinjection.</td>
</tr>
<tr>
<td>Blood</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>to feed mosquitos</td>
</tr>
<tr>
<td>Anopheles stephensi</td>
<td>Animal</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Mosquitos, male and female.</td>
</tr>
<tr>
<td>Drosophila culture vial</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>for egg laying</td>
</tr>
<tr>
<td>Petri dishes</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>with isotonic buffer, for embryo hatching</td>
</tr>
<tr>
<td>Fine paintbrush</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Sable # 0000</td>
</tr>
<tr>
<td>3MM Whatman paper</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Forceps #5</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>double sided tape</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Microloader</td>
<td>&nbsp;</td>
<td>Eppendorf</td>
<td>5242 956.003</td>
<td>&nbsp;</td>
<tr>
<td>Transjector</td>
<td>&nbsp;</td>
<td>Eppendorf</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Microscope</td>
<td>&nbsp;</td>
<td>Leica</td>
<td>&nbsp;</td>
<td>with moving stage and  micromanipulator, set at 10x magnification </td>
</tr>		</tbody>
		</table>
		</div>
			<div>
					</div>
		

injection of an. stephensi embryos to generate malaria-resistant mosquitoes

<p class="jove_content">Die Einführung von exogenen Genen in den Genomen von Moskitos erfordert Mikroinjektion Techniken abgestimmt auf die spezifischen Arten von Interesse. Dieses Video-Protokoll beschreibt eine Methode durch die James-Labor verwendet werden, um DNA-Konstrukte in Anopheles stephensi Embryonen für die Erzeugung von transformierten Mücken microinject. Techniken für die Herstellung Mikroinjektion Nadeln, Sammlung und Aufbereitung von Embryonen und die Durchführung der Mikroinjektion dargestellt.</p>

Watch this Scientific Journal Video about Injection of An. stephensi Embryos to Generate Malaria-resistant Mosquitoes at JoVE.com

Injection of An. stephensi Embryos to Generate Malaria-resistant Mosquitoes

<p class="jove_content">Anopheles stephensi Mücken sind Vektoren für Malaria bewohnen Indien und ganz Asien. Dieses Video zeigt die Technik für die Durchführung microinjections dieser Art mit Transgene, dass Widerstand gegen die Malaria, die Mücke wird zu verleihen. Ein großer Teil der Methodik in diesem Video gezeigt, ist für die Mikroinjektion Techniken anderer Mückenarten.</p>

Injektion eines. stephensi Embryonen zu generieren Malaria-resistenter Moskitos

The introduction of exogenous genes into the genomes of mosquitoes requires microinjection techniques tailored to the specific species of interest. This ...

Research

JoVE Journal

Biology

Microinjection of A. aegypti Embryos to Obtain Transgenic Mosquitoes

Mikroinjektion von A. aegypti Embryonen, um transgene Moskitos erhalten

In this video, Nijole Jasinskiene demonstrates the methodology employed to generate transgenic Aedes aegypti mosquitoes, which are vectors for dengue ...

Forschung

Biologie

In Utero Intraventricular Injection and Electroporation of E16 Rat Embryos

In Utero intraventrikuläre Injektion und Elektroporation von E16 Rattenembryonen

In-utero in-vivo injection and electroporation of the embryonic rat neocortex provides a powerful tool for the manipulation of individual progenitors ...

In Utero Intraventricular Injection and Electroporation of E15 Mouse Embryos

In Utero intraventrikuläre Injektion und Elektroporation von E15 muriner Embryonen

In-utero in-vivo injection and electroporation of the embryonic mouse neocortex provides a powerful tool for the manipulation of individual progenitors ...

Microinjection of Zebrafish Embryos to Analyze Gene Function

Mikroinjektion von Zebrafisch-Embryos, um Gene Function Analyze

One of the advantages of studying zebrafish is the ease and speed of manipulating protein levels in the embryo.  Morpholinos, which are synthetic ...

RNAi Interference by dsRNA Injection into <em>Drosophila</em> Embryos

RNAi Interference by dsRNA Injection into Drosophila Embryos

RNAi-Interferenz durch dsRNA Injektion in<em> Drosophila</em> Embryos

Genetic screening is one of the most powerful methods available for gaining insights into complex biological process 1. Over the years many improvements ...

Imaging <em>C. elegans</em> Embryos using an Epifluorescent Microscope and Open Source Software

Imaging C. elegans Embryos using an Epifluorescent Microscope and Open Source Software

Imaging<em> C. elegans</em> Embryos mit einem Epifluoreszenz-Mikroskop und Open Source Software

Cellular processes, such as chromosome assembly, segregation and cytokinesis,are inherently dynamic. Time-lapse imaging of living cells, using ...

An <em>in vivo</em> Crosslinking Approach to Isolate Protein Complexes From <em>Drosophila</em> Embryos

An in vivo Crosslinking Approach to Isolate Protein Complexes From Drosophila Embryos

Ein<em> In vivo</em> Vernetzung Ansatz zur Proteinkomplexe Von Isolieren<em> Drosophila</em> Embryonen

Many cellular processes are controlled by multisubunit protein complexes. Frequently these complexes form transiently and require native environment to ...

A Multi-detection Assay for Malaria Transmitting Mosquitoes

Ein Multi-Nachweistest für Malaria tragenden Mücken

The Anopheles gambiae species complex includes the major malaria transmitting mosquitoes in Africa. Because these species are of such medical importance, ...

Protocols for Testing the Toxicity of Novel Insecticidal Chemistries to Mosquitoes

Protokolle für die Prüfung der Toxizität von neuartigen Insektizide Chemikalien, Mücken

New classes of insecticides with novel modes of action are needed to control insecticide resistant populations of mosquitoes that transmit diseases such ...

Forced Salivation As a Method to Analyze Vector Competence of Mosquitoes

Gezwungen Speichelfluss als Methode zur Vektorkompetenz von Stechmücken Analyse

Vector competence is defined as the potential of a mosquito species to transmit a mosquito-borne virus (mobovirus) to a vertebrate host. Viable virus ...